Special Issue Preface pubs.acs.org/JPCB
Cite This: J. Phys. Chem. B 2018, 122, 10974−10980
Autobiography of William A. Eaton
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The lives of scientists, considered as Lives, almost always make dull reading. For one thing, the careers of the famous and the merely ordinary fall into much the same pattern, give or take an honorary degree or two, or (in European countries) an honorific order. It could be hardly otherwise. Academics can only seldom lead lives that are spacious or exciting in a worldly sense. ... If a scientist were to cut his ear off, no one would take it as evidence of a heightened sensibility; ...
working toward her Ph.D. in English at Penn. We have been married for 56 years and had two daughters. Elizabeth McBride Eaton graduated from Oldfields School and Wells College with honors in philosophy, was a talented poet, suffered terribly from mental illness, and took her own life at age 38, a tragedy from which we will never recover. Helen Sherman Eaton graduated from Sidwell Friends School and Penn as a French major, from The Juilliard School as violist, earned an M.A. in musicology at the University of Chicago, married the French violinist Guillaume Combet, has two children, and is currently a major figure in the music world in Philadelphia as Chief Executive Officer of the Settlement Music School, the largest community music school in the country. In my youth, I always had to hold a job to have spending moneydelivering newspapers at age 11, working in a laundry at 13, working in a cousin’s photography studio at 15, tutoring algebra and geometry in high school, and performing numerical calculations in college before the age of computers for the research group of the controversial electrochemist, John Bockris. So, I had little time for leisure activities. I was an excellent student in high school, winning the mathematics, physics, and Latin prizes when I graduated from West Philadelphia High School in January 1956 with a 99.6/100 grade-point average and won a full-tuition scholarship to Penn. I was also an excellent student in college and was particularly honored by being the only non-math major in my year to be selected for the honorary mathematics society. I often recall a comment from the math professor, Pincus Schub, who ran the society and frequently reminded us students that “smart people are a dime a dozen; it’s glue on the tuches that counts”. I worked hard at academics, not because I was interested in the subject matterthe only subjects I really liked in college were math, physical chemistry, and Germanbut because I knew that outstanding grades would be necessary to get a scholarship to Penn medical school and, consequently, to enter a profession with a reasonable income that would avoid the financial stress my parents experienced. As an undergraduate, I worked on two very different research projects during the summers, in addition to continuing my year-round, part-time job with Bockris. The first was a repetition of the famous Miller−Urey origin-of-life experiment, with the addition of powdered laevo-rotatory quartz to test the idea of a retired botany professor, Edgar Wherry. Wherry conjectured that proteins are made up of Lamino acids because the original synthesis was catalyzed by an optically active surface. Unfortunately, the chemistry department’s antiquated Rudolph polarimeter prevented me from determining whether the products showed any optical activity. The second was a rather uninteresting project of studying the effects of X-radiation on yeast with a physics professor, Thomas Wood. Toward the end of my senior year, I made one of the best decisions of my life, which was to spend a year in
Peter B. Medawar, New York Review of Books (October 10, 1968)
As a young man growing up in a middle-class neighborhood in West Philadelphia, I had no interest at all in science. Never played with a chemistry set or built anything. As a teenager, I was much more interested in girls, sports, cars, and hanging out with my best friend, Wilfred (“Willy”) Klein. My lifelong passion for science and research did not begin until I started to work with Robin Hochstrasser as a graduate student. I was born on June 4, 1938, in Philadelphia and brought up in a financially poor but intellectually rich family. In spite of very limited means, my parents made sure that all five children received a college educationthree at the University of Pennsylvania and two at Drexel University, both only a mile from our home on South 44th Street. My Jewish mother, Fanny Sherman, daughter of Rabbi Simon Sherman, was born in London, England, in 1898, emigrated to Philadelphia in 1908, was the first in her family to go to college, graduated first in her class from Temple University, received a Master’s Degree in Latin from Penn, and was a high school Latin teacher before she began to raise her five children. My Methodist father, George Washington Eaton, Jr., whose family came to Philadelphia from Wales in the early 18th century, was born in Philadelphia in 1899, joined the Merchant Marines at age 16, made dangerous crossings of the Atlantic during the First World War as a radio officer, married my mother in 1929, lost his electronics business because of the Great Depression, and took a job working for the City of Philadelphia as an electrical technician. He was technically skilled in dealing with anything electrical or mechanical and took courses in electrical engineering at Penn but did not receive a degree. My mother’s Master’s Degree ceremony was held on June 13, 1927. On the very same day, my wife’s father, Thomas D. McBride, graduated from the Law School. He was later to become the legendary Philadelphia lawyer, best known for helping to put an end to the 1950s McCarthy inquisition in Philadelphia when his cross examination of government witnesses led to a charge of perjury during his pro bono defense of eight school teachers accused of being communist subversives. Thirty years later, I met Gertrude McBride when we were both undergraduates at Penn and I often joke that my mother and her father probably looked at each other and smiled that June day in 1927, but it took another round of genetic recombination for the two families to connect. My mother was extremely well read in history and literature and loved discussing these topics with Gertrude, while she was This article not subject to U.S. Copyright. Published 2018 by the American Chemical Society
Special Issue: William A. Eaton Festschrift Published: December 13, 2018 10974
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Berlin before starting medical school as the first “Willy Brandt” exchange student from Penn. To describe what I did in Berlin at this point is too long a digression to present here, so it appears as a separate section at the end. However, I should point out here that I effectively grew up in Berlin from a ̈ Philadelphia boy to a considerably more provincial, naive sophisticated adult. I was not a particularly good student at medical school, except in the course on Biochemistry. The basic science faculty of the medical school strongly encouraged students to do research, in the hope that some of us would pursue a career in research instead of becoming practicing physicians (only two of us became full-time researchers in our class of 120). The medical school also provided summer fellowships just for the asking, even for doing research abroad. In my first summer, I worked in the laboratory of a muscle biochemist, Robert Davies. The experiment I did, which was entirely my idea, showed that phosphocreatine broke down to creatine and phosphate in proportion to the work done in an isotonic contraction of the isolated frog rectus abdominis muscle, thereby answering a long-standing question, albeit indirectly, that ATP is in fact the primary energy donor for muscle contraction. In July 1961, I was fortunate to be at a plenary session of the FASEB meeting in Atlantic City, when Marshall Nirenberg spoke about his experiments to crack the genetic code, followed by Sydney Brenner. I knew almost nothing about this area of science but was enormously impressed with Brenner’s obvious brilliance and incredibly charismatic style. I did not like the muscle field and wanted to try something new for my next research project. So, I wrote to Brenner asking if I could come to his lab for the summer of 1962, with all expenses paid by the medical school. In response, I received a thin aerogram with one line: “Dear Bill, Come if you like, Sydney.” I married Gertrude McBride on June 16, 1962, and started at the new Medical Research Council Laboratory of Molecular Biology (LMB) on Hills Road, Cambridge, just a few days later. My project, supervised by Sydney’s postdoctoral fellow Robin Munro, was to use the in vitro protein biosynthesis system already established in the lab to purify “peptide bond synthesis factor B”. I worked day and night with no success (because it was RNA?). Nevertheless, it was a rewarding experience. For most days that summer, the Brenner group met at least once every day at the canteen on the third floor run by Max Perutz’s wife, Giselacoffee in the morning, lunch, tea at 4 PMusually with Sydney’s pal, Francis Crick. They were an intellectually intimidating duo, so we all just sat quietly and listened to their quite sophisticated discussion, which included defining the important questions of modern biology. It is a pity that I did not know enough molecular biology to more fully appreciate their discussions. However, I did come away that summer with an important lesson about scientific research. Find an important question and, better yet, answer it before anyone else even knows that there is such a question. The next summer I carried out research in the laboratory of Philip George, a well-known hemeprotein physical chemist recruited to Penn from Cambridge by Britton Chance. Working with his postdoctoral fellow, Abel Schejter, I proposed that enzymes do not fold to the thermodynamically most stable state but to the kinetically most accessible state, which is enzymatically active (at the time, I knew nothing about Chris Anfinsen’s work). At each turnover, there is a possibility, albeit with a low probability, that the enzyme will
make a conformational change to the thermodynamically most stable and enzymatically inactive state, and would be recognized as a dead protein by proteolytic enzymes. I thought I had a great new idea about both protein folding and protein regulation. The experiment was to compare the enzymatic activity and susceptibility to proteolysis of two catalase samples, one which was turning over an innumerable number of times converting hydrogen peroxide to oxygen and water and another “resting” without hydrogen peroxide. After one week, there was no difference in either property. In spite of this failure and the one in Cambridge the previous summer, I was committed to pursuing a career in basic research, so I did not want to take an internship. Moreover, the dean, Sam Gurin, advised me not to; otherwise, because of the Vietnam War, I would probably be drafted as a physician to satisfy my selective service obligation. The day after graduation from medical school in June 1964, I felt this enormous weight had just been lifted. I could now study what I wanted for the first time in over a year. I spent the day reading A History of Chemistry by F. J. Moore and W. T. Hall from cover to cover and a few days later went to Philip George’s lab to start serious research as a graduate student supported by a lucrative fellowship from the Pennsylvania Plan to Develop Scientists in Medical Research. Philip and Abel had wanted me to determine the thermodynamic parameters of the cytochrome c redox couple. However, that immediately changed for two reasons. To my great disappointment, Abel was recruited to found and become chair of the first department of biochemistry at the new medical school in Tel-Aviv. In addition, George Hanania, Philip’s former Ph.D. student in Cambridge, had just arrived from Indiana University for a one-year sabbatical. Hanania convinced me that, before performing the difficult potentiometric titration of cytochrome c, I should learn to do potentiometry on a simpler system and nail down the thermodynamic properties of the hexacyanoferrate redox couple, a subject of long-standing interest to both him and Philip. George Hanania was incredibly nice to me and taught me how to do rigorous solution physical chemistry. Beckman had just started to sell potassium ion sensitive electrodes, and I used them to accurately measure the equilibrium constants for binding potassium ions to the hexacyanoferrate anions. In the Fall of 1964, I began graduate courses as part of the Ph.D. program with the relatively new Molecular Biology Graduate Group. I received credit for most of my basic science courses in medical school, so, unlike other graduate programs at Penn, I could spend the majority of my first year of graduate school in research. Because thermodynamics was the virtual religion of Philip’s lab, I pored over thermodynamics texts Bridgman, Callen, Denbigh, Glasstone, Klotz, Lewis and Randall, and Planck. I did audit a graduate course on statistical mechanics that used “Introduction to Statistical Thermodynamics” by Terrell Hill, who later became my tennis partner and one of my closest friends at NIH. In retrospect, instead of studying thermodynamics, I wish I had spent all that time studying statistical mechanics. Statistical mechanics is by far the most important theoretical subject for most biophysical research. Beginning in 1965, I would often meet three young faculty members in the early evening at the Deck Bar at 34th and Walnut StreetsPhilip’s research assistant professor, Alan Adler, Joseph Higgins, a theorist from Chance’s Johnson Foundation, and Robin Hochstrasser, a molecular spectro10975
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National Institutes of Health (NIH) in Bethesda, Maryland. With the help of Gary Felsenfeld, whom Philip knew, I was lucky to land a position with his colleague in the neighboring laboratory in Building 2, Elliot Charney, a spectroscopist interested in optical activity. I departed Philadelphia on January 15, 1968, to become a PHS medical officer at NIH. My colleagues in Robin’s lab felt sorry for me. They thought that with an MD and Ph.D. working with Robin that I would be off to postdoc with one of his famous scientist friends and then on to a career in Academia. Instead, I was headed to a federal research laboratory, completely unknown to them. I could not have asked for a better supervisor. Elliot gave me complete freedom to do whatever I wanted and was always available for help when I needed it. Because of his interest in optical activity, I spent time going into the theory in depth, studying the semiclassical derivation of rotational strength in the quantum chemistry text by Eyring, Walter, and Kimball. With Robin, I was only concerned with the pi electron transitions of the porphyrin ring and charge-transfer bands. The next logical spectroscopic project was to locate the d → d transitions of the iron. I therefore studied crystal field theory, first from elementary texts and slowly graduating to the monumental work of John S. Griffith, “The Theory of Transition Metal Ions”. Crystal field and group theories predict the d → d transitions with large magnetic dipole transition moments, making them more readily detectable in circular dichroism (CD) experiments because of the large ratio of CD to absorption. The Cary CD instrument at NIH could not make measurements at wavelengths longer than about 700 nm, where these transitions were expected, so I went to Cary’s California lab to use their newly developed near-infrared CD instrument for measurements on ferrocytochrome c. Purchasing this very expensive instrument was out of the question. However, I realized that I could make CD measurements with our Cary 14 dual beam absorption spectrophotometer by inserting a polaroid in both compartments to obtain linear polarization followed by plastic quarter wave retarders to create left- and right-circularly polarized light and measure CD as the difference in optical density. I used my $20 CD attachment to discover the near-infrared d → d transitions of the iron−sulfur proteins, rubredoxin, ferrodoxin, and adrenodoxin, to show that the iron in these proteins of unknown structure is tetrahedrally coordinated. After about one year, Elliot offered me a permanent civil service position. I arrogantly and foolishly dismissed the idea, thinking that after satisfying my two-year selective service obligation in January 1970 that I would be moving on to a faculty position at a major research university. However, a recession arrived in 1969 and very few universities were hiring. I was quite depressed about my situation, when a conversation with a close friend of Elliot’s, Marty Gellert in the neighboring Laboratory of Molecular Biology, completely changed my attitude about NIH. I was complaining to Marty, when he asked: “Why do you want an academic job? This is the best place in the country to do research and you told me you liked the life of a research scientist at the LMB in Cambridge.” It hit me like a ton of bricks. It was not until several years later that my fellow graduate students, who by then realized the greatness of the NIH, would remind me that I had landed the best job in all of science. In fact, during my 50 years at NIH, I only left for a significant period of time once. That resulted when I met Wally Gilbert at a Biopolymers Gordon Conference in 1974, where I
scopist in the chemistry department. Listening to the three argue vehemently over what was the most important theoretical discipline for chemistry was both educational and enormous fun for me. Alan was wedded to thermodynamics, Joe to statistical mechanics, and Robin to quantum mechanics. One evening I learned that Robin’s research focused on the optical properties of molecular crystals. I told him about a conformationally sensitive, near-infrared optical absorption band of cytochrome c at 695 nm and a huge crystal that Emanuel Margoliash, Abel Schejter’s Ph.D. supervisor and the dean of cytochrome c research, had given me. He immediately demanded that I fetch the crystal and bring it to his lab, even though it was close to midnight and neither of us was completely sober. Robin quickly jerry-rigged an optical system with a microscope and collected a polarized absorption spectrum in the near-infrared of the almost black crystal with a 3 m spectrograph. After a few minutes, he emerged from the dark room to announce with great excitement that “your 695 band is z-polarized.” I had not the foggiest idea what he was talking about. Robin then gave me a short, very enthusiastic lecture on the importance of transition moment directions in interpreting the quantum mechanical origin of electronic transitions, but I still did not quite understand. Two weeks later, he presented this work at the 1966 international meeting on hemeproteins at the Johnson Foundation. I was bowled over by his talk describing the 695 band result, which also greatly impressed the many cytochrome aficionados in attendance. Robin made research in molecular spectroscopy sound so exciting that I decided I had to work with this guy. Moreover, he told me with great certainty that I could make hemeprotein single crystal spectroscopy a great Ph.D. project and advised me to abandon solution physical chemistry, which he called “stone age science”. I spent the next 18 months with Robin, where, with the help of his very kind postdoc, George Castro, I put together a microspectrophotometer to measure polarized spectra of sperm whale myoglobin and several cytochrome c crystals from different species, including horse, tuna, duck, and snapping turtle. The instrument I assembled consisted of a Leitz polarizing microscope, a Leitz photometric attachment powered by three, 300 V batteries, a xenon lamp, a grating plus prism double monochromator, and a field diaphragm and pinhole to provide the double confocal masking necessary for measuring high optical densities. Robin gave me many long private tutorials in molecular spectroscopy, group theory, quantum mechanics, and molecular orbital theory. I reciprocated somewhat by educating him in biochemistry. Robin was not only responsible for changing my interest in scientific research into a genuine passion but became my life-long best friend until he died on February 27, 2013. My life changed dramatically in November 1967 when I received a draft notice in the mail. I immediately rushed to the selective service office in Philadelphia to explain that I was not a licensed physician and, therefore, was of no use to the military. Dean Gurin was dead wrong. An at first seemingly mean-spirited bureaucrat opened up a thick book on selective service regulations and asked me: “Dr. Eaton, do you have a doctor of medicine degree?” When I replied “yes”, she pointed to a section in the book and said “you are drafted”. She was, however, extremely helpful by telling me about an organization of which I had never heardthe Commissioned Corps of United States Public Health Service (PHS)and that Commissioned Officers could carry out research at the 10976
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their meeting and pay all expenses to hear a talk about their mistake? Since I was a last-minute invitee, I was not on the program, so I was only given time to show one slide by the session chair. It was my first international meeting as an independent investigator, and I was very nervous about giving the talk in front of so many eminent scientists, which, in addition to Perutz, included hemoglobin luminaries of the day such as Eraldo Antonini, Maurizio Brunori, Quentin Gibson, John Hopfield, Martin Karplus, Robert Shulman, and Jeffries Wyman. I was, nevertheless, able to make my point clearly with a single slide, because I practiced what I would say over and over again through a sleepless night. What followed was the first major turning point in my career. It was now 1974, and I realized that molecular spectroscopy was not a field where I was going to thrive. Interpreting optical spectra from different kinds of measurements and quantum chemical theoretical methods was often both interesting and intellectually challenging, but it started to seem like accountant’s work. I decided that I wanted to learn more about sickle hemoglobin aggregation. Elliot was incredibly generous by also allowing Jim to work with me on this project. Our first experiment was to induce the formation of a deoxyhemoglobin S gel by raising the temperature from 0 to 20 °C. Nothing happened for about 20 min. Then, there was a sudden appearance of very bright birefringence; i.e., there was a long lag phase before the appearance of the gel, which we called a delay period. We then lowered the temperature back down to 2 °C and observed that the birefringence disappeared without a delay in just a couple of minutes. Repeating this experiment several times at several different elevated temperatures showed that sickle hemoglobin gelation was extremely temperature-sensitive, with an Arrhenius plot of the duration of the delay period, which we called a delay time, versus the reciprocal absolute temperature to give a slope corresponding to a 90 kcal/mol activation energy. Philip Ross from the neighboring Laboratory of Molecular Biology became interested and joined forces with us to use differential scanning calorimetry to make kinetic as well as thermodynamic measurements. More remarkable than our finding of a very large activation energy was our discovery of the unheard of enormous sensitivity of the delay time to concentration, where we determined that the reciprocal of the delay time depends on the 30th power of the initial sickle hemoglobin concentration. Then came my “eureka moment” in scientific research, where 4 years of medical school really paid off. In a very short period, I recognized the medical relevance of our findings. First, the delay period could prevent obstruction of the circulation in tissues by allowing many or even most cells to escape the capillaries before the cells sickle. Consequently, the relation between the delay time and the capillary transit time could be the primary determinant of clinical severity. Furthermore, it would not be necessary to completely inhibit sickle hemoglobin polymerization to treat sickle cell disease. Simply increasing the delay time to allow more cells to escape the capillaries before sickling could be therapeutic, and this could be achieved from even a very small decrease in the intracellular sickle hemoglobin concentration. For the next 12 years, Jim and I elaborated on this initial work with an extensive series of thermodynamic and kinetic studies and development of a novel nucleation mechanism, with the collaboration of three wonderful postdoctoral fellows, Helen
presented our initial work on sickle hemoglobin aggregation, described below. Wally invited me to Harvard to give a biochemistry seminar in the Fall of 1974. I was then asked to teach the senior/first year graduate course in physical biochemistry with Steve Harrison for the Spring semester of 1976 as a Visiting Professor of Biochemistry. Steve taught elementary statistical mechanics and structural biology, and I taught elementary quantum mechanics and kinetics. At the end of the semester, I gave a physical chemistry seminar, which apparently impressed the faculty. For in the Fall, Arthur Solomon, who was retiring as chair of the Biophysics Graduate Group, called me to tell me that I was the unanimous choice of the biophysics faculty to replace him, which came with a tenured faculty appointment in the chemistry department. The next day, before I even had any chance to think about whether or not I would leave the NIH, Solomon called back to tell me that he had to withdraw the offer because the appointment in chemistry was approved by the physical chemists without consulting the organic chemists. Back to 1969. My next project was to assemble a microspectrophotometer very similar to the one in Philadelphia but with precious, chromatically corrected, strain-free Zeiss (“ultrafluar”) objectives. After determining transition moment directions of nucleic acid bases from polarized UV single crystal spectra with these objectives, I returned to hemeproteins. I collaborated with Marvin Makinen, who made beautiful hemoglobin crystals for polarized optical absorption measurements. It was these spectroscopic studies on hemoglobin crystals that aroused my interest in sickle cell disease research, a research area that was exploding in the early 1970s. I had learned that a colleague in my institute, Makio Murayama, had concluded in a 1966 paper in Science that the hemes are parallel to the long axis of the fibers inside sickled cells. So, I decided to determine the orientation of the hemes in sickled cells unambiguously by using polarized absorption. In the course of my measurements, Jim Hofrichter had just arrived as a postdoctoral fellow to work with Elliot. Jim became interested in my project and with his great technical skills immediately made several improvements to my instrument. Our measurements showed that the heme planes are nearly perpendicular to the long axis of sickled cells, so that the long, pseudo-2-fold axis of the hemoglobin molecule, designated as the x axis, must be less than 22° from the long axis of the sickle fibers inside the cells. We did not realize the importance of these measurements until learning about a fiber structure just published in PNAS by John Finch and Max Perutz in Cambridge. Jim and I went to Richard Feldman at the computer division of the NIH to investigate the relation between our result and this proposed structure for the sickle fibers. Richard was a true pioneer in molecular graphics, and we were able to examine the hemoglobin structure in atomic detail. While our optical result was compatible with the Finch/ Perutz structure, we discovered on Richard’s graphics that the true 2-fold axis of the hemoglobin molecule, which was pointing radially from the fiber in their structure, did not permit the site of the beta6 glutamic acid to valine mutation to be an intermolecular contact in the fiber. I wrote to Perutz telling him about our result and what I believed was a mistake by him. He immediately wrote back inviting me to present our results at his hemoglobin meeting at the Royal Society in January 1973 and to also pay for all of my travel and hotel expenses. How many scientists today would invite someone to 10977
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folding”. I focused most of my efforts on protein folding from 1993 to 2015, with many of my studies motivated by Peter’s pioneering theoretical work. I will not say more about my protein folding studies or the single molecule work that started with Ben Schuler and Everett Lipman and continued with my now colleague Hoi Sung Chung. Instead, I would like to mention work that was particularly exciting to me but not to many others, as judged by their low citation record. Some of my favorites that have been poorly cited are (i) derivation of kinetic equations, which showed that a large part of the concentration dependence of the delay time in nucleated polymerization can result from the concentration derivatives of the activity coefficients, (ii) discovery of a relation between hemoglobin exons and functional parts of the molecule, (iii) demonstration with Andrea Mozzarelli that the T quaternary structure of hemoglobin binds oxygen noncooperatively in single crystal experiments, confirming the major postulate of the two-state allosteric model of Monod, Wyman, and Changeux (MWC) and thereby settling a long-standing controversy, (iv) demonstration of a linear free energy relation for the quaternary conformational transition of hemoglobin and suggesting the position of the transition state along the reaction coordinate from the exponent relating rate coefficients and equilibrium constants, (v) measurement of distributed ligand binding kinetics to myoglobin at room temperature that look like its low temperature distributed kinetics by immersing the protein in a trehalose glass, (vi) determination of the distance dependence of the quenching rates of the tryptophan triplet by cysteine by measuring the kinetics in a trehalose glass dilute in tryptophan and concentrated in cysteine, (vii) realization that a very unusual result of Andrea and Cristiano Viappiani in Parma on carbon monoxide rebinding to hemoglobin after photolysis in silica gels could be explained by our extension of the MWC model to include tertiary conformational equilibria, and (viii) my retrospective analysis of Pauling’s famous 1949 sickle cell paper in Science and his effective retraction 5 months later. The most recent turning point came in 2006 when Dan Camerini, a molecular biologist in a neighboring lab in Building 5, convinced me to return to sickle cell research. Hydroxyurea was still the only drug approved by the Food and Drug Administration for the treatment of the disease, and it was only partially successful. Dan’s prompt had a big effect on me. I started what was at first a part-time drug screening project while I worked primarily on single molecule protein folding experiments. The project was initiated with an instrument and image analysis developed by a brilliant postdoctoral fellow, Jeffrey Smith, a Ph.D. student of Chris Dobson. About 3 years ago, I started working almost full-time on this project. In fact, the plan for the rest of my research career is to try to discover drugs in addition to hydroxyurea by developing sensitive, high throughput screening assays and applying them to libraries of compounds, especially those already tested in humans. It is very high risk but also very high payoff research. Finally, I should say that whatever success I have had in science resulted from three major factors. First and most importantly has been my good fortune of having many very talented postdocs and brilliant senior co-workers in Jim Hofrichter and Eric Henry, and an incredibly smart and enormously generous theoretical colleague, Attila Szabo. Attila has always been willing to immediately stop whatever he was
Sunshine, who had a very successful career in scientific administration at NIGMS/NIH, Frank Ferrone, a Ph.D. student of John Hopfield at Princeton, who was key in formulating the double nucleation mechanism, and has continued to do beautiful work on sickle hemoglobin aggregation to this day in the physics department at Drexel University, and Andrea Mozzarelli, a junior co-worker of my dear friend Gian Luigi Rossi at the University of Parma, who performed technically demanding experiments to show that the delay period prevents most cells from sickling in vivo, indicating that patients survive sickle cell disease because of the kinetics. Andrea and I have collaborated on studies of hemoglobin allostery on and off for the past 28 years. It was not easy to convince hematologists of the importance of our work, primarily because they thought that physical chemistry could not possibly explain what was going on in a complex human disease. Many would approach me after talks to tell me that what I said was certainly wrong because it was inconsistent with their experience in treating one of their patients. H. Franklin Bunn, a brilliant hematologist and biochemist at Harvard Medical School, widely considered as the world’s leading expert on hemoglobinopathies, came to our rescue by clearly explaining the importance of our work to the medical community. Frank has not only always given me help and great advice on all things clinical and biochemical, but he has been a dear friend to me and my family for over 40 years. The next major turning point in my career started on October 24, 1994, when I woke up totally refreshed at the Johns Hopkins Bayview Hospital after only 4 hours of sleep but with continuous positive airway pressure for the first time. Gertrude first recognized that I was not breathing properly at night after surgery in 1983, so my brain had been functioning at a relatively lower level for 11 years, ironically due to decreased arterial oxygenation of my hemoglobin from sleep apnea. That day was the beginning of an enormous increase in my physical energy, as well as an increase in scientific productivity in the field of protein folding. The protein folding work was motivated by a conversation over vodka with the famous theorist, Peter Wolynes, at a meeting in Moscow in June of 1991. At that time, I had been working on nanosecond-resolved kinetics of ligand rebinding and conformational changes in hemoglobin and myoglobin following carbon monoxide photodissociation, still collaborating with Jim Hofrichter, who constructed a high precision, nanosecond resolved pump and probe spectrometer. My interest in measuring transient spectra in fast kinetic studies was motivated by an earlier collaboration with Robin on picosecond-resolved spectra of hemoglobin. Another very smart student of John Hopfield, Eric Henry, arrived in 1980, who was tremendously skilled at computing and developed elegant methods for analyzing the time course of Jim’s transient spectra using singular value decomposition. Peter suggested that I use my lab’s fast kinetic methods to work on a “difficult problem”. The difficult problem was protein folding. I did not do anything until a visit from Heiner Roder. Heiner told me that we could use nanosecond laser pulses to initiate folding of chemically denatured ferrocytochrome c by photodissociating its carbon monoxide complex. It was a perfect experiment for my labJim’s instrument, Eric’s sophisticated analysis, carbon monoxide photolysis, and my favorite molecule from graduate school. The experiment improved the time resolution in protein folding experiments by almost 6 orders of magnitude and started the subfield of “fast protein 10978
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doing to spend as much time as I wanted to help me whenever I got stuck with any problem, theoretical or experimental. The second factor, although not as important as the first, has been my ability to ask important questions and my persistence in answering them (Pincus Schub’s “glue on the tuches”), together with my obsessive compulsiveness in paying attention to the tiniest of details in both performing and supervising experiments. Finally, I have been fortunate to have worked in an extraordinarily stimulating and supportive scientific environment at NIH, surrounded by outstanding colleagues in the Laboratory of Chemical Physics, every one of whom is currently considered as among the world leaders in his area of research.
Agency (ASA), translating Russian from radio communications of Soviet troops surrounding Berlin from US Army listening posts at Tempelhof Airport. Marty was obviously concerned, and the next day, he charged into our apartment white-faced to tell me that Leonid Dmitrievich Mozgin was in fact the head of the KGB in East Berlin and that he (Marty) would suffer severe consequences or possibly even face a court-martial for illegally living off base with a roommate who was in contact with the KGB. The only way to prevent this was for me to fully cooperate with the Army Counter Intelligence Corps (CIC) that watched over the ASA. The next day, standing at the bus stop at the corner of Riemeister Strasse and Sophiecharlotte Strasse, where I was to meet with someone from the CIC, a black sedan pulled up to the bus stop, and a man who was also at the bus stop reading a newspaper, pushed me into the back seat of the sedanit seemed like something from a Hollywood spy movie. We drove to a café next to the Wannsee, the largest lake of Berlin, where the two CIC agents gave me a patriotic speech about how I could help my country by meeting with Mozgin and recounting to them exactly what he said. I agreed, but at this point, I realized I was in over my head and expressed fear of meeting Mozgin even though it was West Berlin. They told me not to worry because they would also be at the café to protect me. The one hour conversation with Mozgin was rather innocuous. I was told by the CIC guys that the conversation was just the beginning of a probe to see if I would be a candidate for defection from the “decadent West” to the communist East, which would have been a great propaganda coup for the Russians. Berlin was crawling with spies in those days. On one occasion, I was approached by a guy in typical ivy league dress outside the Staatsoper, who tried to befriend me. He spoke perfect English with no accent and knew a lot about baseball. Marty Rice was with me, and once we were inside the opera house, Marty heard the guy in one of the stalls in the men’s room speaking perfect Russian. A particularly memorable occasion was when the CIC needed to know if Mozgin was in Berlin, so they wanted me to go to East Berlin and telephone the embassy. They drove me to the crossing point at Potsdamer Platz. I told them that I was hesitant to call the embassy from the public phone booth just across the border with so many armed East German border police. They assured me that I would be safe and if any trouble did arise the guy in the back seat, whom I had never met before, was a CIC sharpshooter. I only saw Mozgin one or two more times. I believe he lost interest in me because it must have been clear to him that I was too much of an American chauvinist to ever be a candidate for defection to the East. Nevertheless, during my last 7 months in Berlin, I spent a lot of time downright scared. In my last meeting with the CIC in Berlin, I was asked about the next Willy Brandt exchange student. I told them his name was Marvin Makinen, but I did not know anything else about him. A month after I returned to Philadelphia, I was asked to meet with the FBI and gave a one hour taped account of my experience with the KGB and CIC guys. I never spoke about any of these experiences to anyone except Gertrude and Marvin for over 40 years. In 2007, I used the Freedom of Information Act to obtain a transcript of the FBI interview for my family and to refresh my memory about everything that happened in Berlin. After several months, I received 65 pages, almost all of them redacted with black ink. One of the few unredacted lines concerned the interview in a memo to the
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MY INTERACTION IN BERLIN WITH THE ARMY SECURITY AGENCY (ASA), THE COUNTER INTELLIGENCE CORPS (CIC), AND THE COMMITTEE FOR STATE SECURITY (KGB) In September 1959, I sailed from New York to Le Havre on the SS United States. On my second day in Berlin, I had lunch with Willy Brandt, then the mayor of Berlin, and other American students at the City Hall in Schoneberg. Berlin was the center of world politics at the time and was such a fascinating city that I barely went to any classes after the first few weeks. I was burned out from working so hard at Penn as an undergraduate that I had no interest in studying the material for advanced courses in mathematics and chemistry from textbooks and lectures in German. I only glanced at physics texts that Thomas Wood gave me in the hope that I would study them and return to graduate school in physics instead of medical school. Instead, I spent most of the time in cafés, bars, and nightclubs discussing politics with German students and going to concerts and operas in East Berlina ticket in what was formerly Hitler’s box at the Staatsoper cost only one US dollar. I was treated extremely well by the Berliners, who were grateful to the Americans for saving the city with an airlift in 1948 and liked us American students who spoke German with correct grammar. I also traveled frequently to visit other major European cities. My life in Berlin changed after a visit in December 1959 with two college friends, Arnold Fisher and Gary Goldschneider, to the Soviet Embassy in East Berlin to find out what we had to do to visit Russia. We met Leonid Mozgin, who introduced himself as the cultural secretary of the embassy. He disabused us of the idea of traveling to Russia, when he told us of the enormous cost. As we left the embassy, Mozgin gave me his calling card and remarked: “Mr. Eaton, I hope we will see each other again, so please give me your telephone number.” Little did I suspect what was going to happen. About 3 weeks later, Mozgin telephoned me. The call came via Switzerland, since there were no telephone lines between East and West Berlin. Mozgin told me that he was becoming interested in Abraham Lincoln but could not find any biographies of him in East Berlin, so would I bring one to him at the embassy. I was leery of the whole idea and said that I would get the book for him from the university library but preferred to meet in West Berlin, to which he immediately snapped: “Fine, Schultheiss Café on the Kurfurstendamm, Tuesday at 1 PM.” I immediately told my part-time roommate in my basement apartment at Sophiecharlotte Strasse 33a in Dahlem, Marty Rice, about the telephone call. I knew Marty from West Philadelphia High School. He was in the Army Security 10979
DOI: 10.1021/acs.jpcb.8b06737 J. Phys. Chem. B 2018, 122, 10974−10980
The Journal of Physical Chemistry B
Special Issue Preface
Director of the FBI, John Edgar Hoover, and said: “During this interview EATON appeared to be slightly effeminate in his speech and in some of his mannerisms.” Another memo, this one from John Edgar Hoover, states that “It is requested that you [name redacted] advise this Bureau if any additional data has been ascertained regarding the subject’s activities in Germany or his association with a member of the Soviet Intelligence Service”. These were the days when being gay could be ruinous to one’s career and the FBI was clearly concerned about the possibility that I could be working for the KGB under the threat of blackmail. In September 1961, I read in the Philadelphia Evening Bulletin that Marvin Makinen was arrested by the Soviets for taking pictures of military installations in the Ukraine (see “A Chemistry Spy Story”, Chemical and Engineering News, 2013, 91, 47−49; https://cen.acs.org/articles/91/i7/Chemistry-SpyStory.html). After 28 months in prison, Marvin was released in a prisoner exchange and entered Penn Medical School. Marvin then came to the NIH as a PHS medical officer, where we worked closely together on single crystal spectroscopy of hemoglobin discussed earlier. We also socialized and drank a lot together, but Marvin never admitted spying. That is, until one evening in 1993 sipping cocktails at a joint 80th birthday celebration of Britton Chance and Mildred Cohn at Brit’s yacht club on the Delaware, when I asked him why he never admitted to me that he was spying. He answered: “because I was caught and you weren’t”.
William A. Eaton
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DOI: 10.1021/acs.jpcb.8b06737 J. Phys. Chem. B 2018, 122, 10974−10980
